摘要
目的：研究人参皂苷Rh2对肝癌Hep G2细胞迁移的影响及其机制。方法：取对数生长期Hep G2细胞,Rh2（10~160μmol/L）诱导,同时设空白对照组,用CCK-8检测Rh2对细胞增殖影响;Transwell检测Rh2对细胞的迁移的影响;荧光素报告基因筛选可能的信号通路;Western blot检测Hep G2细胞中P-ERK、ERK、P-P38、P-38、P-JUK、JUK、MMP3等蛋白的表达;定量PCR方法检测AP1、MMP3基因的表达;荧光显微镜分别观察AP1、MMP3蛋白在细胞内的表达及分布。结果：CCK-8检测结果显示Rh2对肝癌Hep G2细胞生长抑制呈剂量和时间依赖性;Transwell检测显示Rh2对肝癌Hep G2细胞迁移有明显的抑制作用;荧光素报告基因筛选出AP1转录因子;Western blot检测结果显示P-ERK、MMP3蛋白表达水平呈下降趋势,PJUK、P-P38蛋白表达水平明显升高,ERK、JUK、P38蛋白则无明显变化;定量PCR方法结果显示：AP1、MMP3的基因表达下降。荧光结果显示：AP1、MMP3均分布在胞质内,随药物浓度的增加其荧光表达量减弱。结论：人参皂苷Rh2可通过激活MAPK通路来抑制肝癌Hep G2细胞的迁移。
Objective： To study the mechanism of ginsenoside Rh2 inhibiting Hep G2 cells migration. Methods： Hep G2 cells in logarithmic growth phase were cultured in 96-well plates,which were induced by different concentration Rh2,respectively for 24,48,72 hours. The cell inhibition was detected by Cell Counting Kit. Transwell chambers was used to checked Hep G2 cell migration ability; luciferase was tested by Luciferase Reporter Assay system reagent; The expressions of P-ERK,ERK,P-P38,P-38,P-JUK,JUK,MMP3 proteins were detect by Western blot; the expression of AP1,MMP3 gene were detected by Quantitative PCR; The expression of AP1,MMP3 fluorescence protein were observed by fluorescence microscopy. Results： Administrated with different concentration of Rh2 after24,48,72 h,the proliferation of Hep G2 cells were inhibited（ P〈 0. 05）,and in dose-and time-dependent manner. Transwell assay showed Rh2 could significantly inhibited migration of Hep G2 cells. The expressions of P-ERK,MMP3 proteins were significantly decreased,the expressions of P-JUK,P-P38 proteins were significantly increased,expression levels of ERK,P-38,JUK were no significant difference. Expression of AP1,MMP3 gene were significantly decreased,the expressions of AP1,MMP3 fluorescence proteins were significantly decreased. Conclusion： Ginsenoside Rh2 can activate MAPK pathway to inhibit the migration of Hep G2 cells.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2015年第1期61-65,共5页
Chinese Journal of Immunology
